Yoshiyuki Kageyama

Population study of sizes and components of self-reproducing giant multilamellar vesicles.

Population analysis of a system of self-reproducing giant multilamellar vesicles (GMVs) was carried out by means of flow cytometry. The multidimensional distribution of forward light scattering (FS), side light scattering (SS), and fluorescence (FL) intensities originating from each GMV provided information about changes in a population composed of 104 vesicles. FS-FL dot plots indicated that, after the addition of the membrane precursor, the size distribution of the newly generated vesicles was nearly the same as that of the original, but the catalyst content was reduced. This result can be interpreted as evidence for the occurrence of the self-reproduction of GMVs. Moreover, the new GMVs recovered the amount of catalyst to the initial value, keeping their size distribution constant, when a solution of the catalyst was added to the new GMVs. These results are the first experimental evidence for a novel phenomenon on GMV size distribution during their self-reproducing cycle.

Design and synthesis of a photocleavable biotin-linker for the photoisolation of ligand–receptor complexes based on the photolysis of 8-quinolinyl sulfonates in aqueous solution

The ability of avidin (Avn) to form strong complex with biotin (Btn) is frequently used in the detection and isolation of biomolecules in biochemical, analytical, and medicinal research. The fact that the binding is nealy irreversible, however, constitutes a drawback in term of the isolation and purification of intact biomolecules. We recently found that 8-quinolinyl esters of aromatic or aliphatic sulfonic acids undergo photolysis when irradiated at 300-330 nm in aqueous solution at neutral pH. In this work, a biotin-dopamine (BD) conjugate containing a photocleavable 8-quinolinyl benzenesulfonate (QB) linker, BDQB, was designed and synthesized for use in the efficient recovery of dopamine-protein (e.g., antibody) complexes from an Avn-Btn system. The complexation of BDQB with a primary anti-dopamine antibody (anti-dopamine IgG(1) from mouse) on an Avn-coated plate was confirmed by an enzyme-linked immunosorbent assay (ELISA) utilizing a secondary antibody (anti-IgG(1) antibody) conjugated with horseradish peroxidase (HRP). Upon the photoirradiation (at 313 nm) of the BDQB-IgG(1) complex, the release of dopamine-IgG(1) complex was confirmed by ELISA. Characterization of the resulting photoreleased dopamine-anti-dopamine IgG(1) complex was performed by SDS-PAGE and Western blot.

Dissipative and Autonomous Square‐Wave Self‐Oscillation of a Macroscopic Hybrid Self‐Assembly under Continuous Light Irradiation

Building a bottom-up supramolecular system to perform continuously autonomous motions will pave the way for the next generation of biomimetic mechanical systems. In biological systems, hierarchical molecular synchronization underlies the generation of spatio-temporal patterns with dissipative structures. However, it remains difficult to build such self-organized working objects via artificial techniques. Herein, we show the first example of a square-wave limit-cycle self-oscillatory motion of a noncovalent assembly of oleic acid and an azobenzene derivative. The assembly steadily flips under continuous blue-light irradiation. Mechanical self-oscillation is established by successively alternating photoisomerization processes and multi-stable phase transitions. These results offer a fundamental strategy for creating a supramolecular motor that works progressively under the operation of molecule-based machines.

Macroscopic motion of supramolecular assemblies actuated by photoisomerization of azobenzene derivatives

Submillimetre size self-assemblies composed of oleate and azobenzene derivatives show forceful motions such as screw-type coiling-recoiling motion by photoirradiation.

Photolysis of the sulfonamide bond of metal complexes of N-dansyl-1,4,7,10-tetraazacyclododecane in aqueous solution: a mechanistic study and application to the photorepair of cis,syn-cyclobutane thymine photodimer.

Sulfonamide constitutes a ubiquitous functional group that is frequently used in organic chemistry, analytical chemistry, and medicinal chemistry. We report herein on the photolysis of a dansylamide moiety of 1-dansyl-1,4,7,10-tetraazzacyclododecane (N-dansylcyclen, L(2)) in the presence of a zinc(II) ion in aqueous solution. By potentiometric pH titrations, the complexation constant for the 1:1 complex of L(2) and Zn(2+), log K(s)(ZnL(2)), in aqueous solution at 25 degrees C with I = 0.1 (NaNO(3)) was determined to be 6.5+/-0.1. The structure of the ZnL(2) complex was confirmed by single-crystal X-ray diffraction analysis. During fluorescence titrations of L(2) with Zn(2+) (irradiation at 308 or 350 nm) in aqueous solution at pH 7.4 (10 mM HEPES with I = 0.1 (NaNO(3))) and 25 degrees C, considerable enhancement in fluorescence emission of the Zn(2+) complex of L(2) (ZnL(2)) was observed, while metal-free L(2) exhibited only a negligible emission change upon UV irradiation. It was revealed that this emission enhancement arose from the photoinduced cleavage of a sulfonylamide moiety in ZnL(2), yielding the Zn(2+)-cyclen complex and 5-dimethylaminonaphthalene-1-sulfinic acid, which has a greater quantum yield (Phi) for fluorescence emission than that of L(2) and ZnL(2). For comparison, the photolysis of N-(1-naphthalenesulfonyl)cyclen (L(3)) and its Zn(2+) complex (ZnL(3)) under the same conditions (irradiation at 313 nm) gave the corresponding sulfonate (1-naphthylsulfonate). We also describe the results of a photoreversion reaction of cis,syn-cyclobutane thymine photodimer (T[c,s]T) utilizing the photolysis of ZnL(2) and ZnL(3).

Compartment size dependence of performance of polymerase chain reaction inside giant vesicles

The amplification of a 1229 bp template DNA that encoded the green fluorescent protein (GFP) was conducted in cell-sized giant vesicles (GVs, ϕ > 1 µm) using a real-time polymerase chain reaction (PCR) technique. The proportion of PCR-proceeded GV reached up to 16% of all GVs. The dependence of PCR on GV size was elucidated by flow cytometry analysis.

Formation of Monodisperse Hierarchical Lipid Particles Utilizing Microfluidic Droplets in a Nonequilibrium State

A new microfluidic process was used to generate unique micrometer-sized hierarchical lipid particles having spherical lipid-core and multilamellar-shell structures. The process includes three steps: (1) formation of monodisperse droplets in a nonequilibrium state at a microchannel confluence, using a phospholipid-containing water-soluble organic solvent as the dispersed phase and water as the continuous phase; (2) dissolution of the organic solvent of the droplet into the continuous phase and concentration of the lipid molecules; and (3) reconstitution of multilamellar lipid membranes and simultaneous formation of a lipid core. We demonstrated control of the lipid particle size by the process conditions and characterized the obtained particles by transmission electron microscopy and microbeam small-angle X-ray scattering analysis. In addition, we prepared various types of core-shell and core-core-shell particles incorporating hydrophobic/hydrophilic compounds, showing the applicability of the presented process to the production of drug-encapsulating lipid particles.

Study on structural changes in supramolecular assemblies composed of amphiphilic nicotinamide and its dihydronicotinamide derivative by flow cytometry

Flow cytometric analysis revealed that the redistribution of components of a hybrid vesicular system composed of amphiphilic nicotinamide and its reduced form occurred associated with the emergence of nonlamellar aggregates that emitted an enhanced blue emission together with a green emission due to exciplex formation.

Mechanism of Macroscopic Motion of Oleate Helical Assemblies: Cooperative Deprotonation of Carboxyl Groups Triggered by Photoisomerization of Azobenzene Derivatives

Macroscopic and spatially ordered motions of self-assemblies composed of oleic acid and a small amount of an azobenzene derivative, induced by azobenzene photoisomerization, was previously reported. However, the mechanism of the generation of submillimeter-scale motions by the nanosized structural transition of azobenzene was not clarified. Herein, an underlying mechanism of the motions is proposed in which deprotonation of carboxyl groups in cooperation with azobenzene photoisomerization causes a morphological transition of the self-assembly, which in turn results in macroscopic forceful dynamics. The photoinduced deprotonation was investigated by potentiometric pH titration and FTIR spectroscopy. The concept of hierarchical molecular interaction generating macroscale function is expected to promote the next stage of supramolecular chemistry.